Noordhoek G T, Kaan J A, Mulder S, Wilke H, Kolk A H
Public Health Laboratory Friesland, Leeuwarden, The Netherlands.
J Clin Pathol. 1995 Sep;48(9):810-4. doi: 10.1136/jcp.48.9.810.
To investigate the use of the polymerase chain reaction (PCR) in the routine laboratory for the detection of Mycobacterium tuberculosis in clinical samples.
Samples were divided and processed separately for the detection of M tuberculosis by microscopy, culture and PCR. After DNA extraction, PCR was performed with primers specific for the insertion element IS6110 and the product was analysed by agarose gel electrophoresis, Southern blotting or dot blotting and hybridisation with a digoxigenin labelled internal probe. Each sample was tested for inhibitors of Taq polymerase with the aid of an internal control. Multiple negative and positive controls were used to monitor each step of the procedure.
The data from two laboratories, using the same operating procedures, were combined. Of 1957 specimens, 79 (4%) were culture and PCR positive, while 1839 (93.9%) were negative in both tests. Thirty specimens (1.5%) were PCR positive only and nine (0.5%) were culture positive but PCR negative.
Using culture and clinical history as the gold standard, sensitivity and specificity for PCR were 92.1% and 99.8%, respectively. With elaborate precautions, PCR is a suitable and reliable method for the detection of M tuberculosis in clinical samples in a routine microbiology laboratory.
研究聚合酶链反应（PCR）在常规实验室中用于检测临床样本中结核分枝杆菌的应用。
将样本分开处理，通过显微镜检查、培养和PCR检测结核分枝杆菌。提取DNA后，使用针对插入元件IS6110的引物进行PCR，产物通过琼脂糖凝胶电泳、Southern印迹或斑点印迹以及与地高辛标记的内部探针杂交进行分析。借助内部对照对每个样本进行Taq聚合酶抑制剂检测。使用多个阴性和阳性对照监测该过程的每个步骤。
合并了两个采用相同操作程序的实验室的数据。在1957份标本中，79份（4%）培养和PCR均为阳性，而1839份（93.9%）两项检测均为阴性。30份标本（1.5%）仅PCR为阳性，9份（0.5%）培养为阳性但PCR为阴性。
以培养和临床病史作为金标准，PCR的敏感性和特异性分别为92.1%和99.8%。采取精心的预防措施后，PCR是常规微生物实验室检测临床样本中结核分枝杆菌的一种合适且可靠的方法。
